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Highly sensitive and facile microRNA detection based on target triggered exponential rolling-circle amplification
Shiying Zhou1, Human Sun1, Jiangbo Dong1
1Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR China.
Analytica Chimica Acta
|May 25, 2023
Summary
We developed a target triggered exponential rolling-circle amplification coupled with CRISPR/Cas12a (T-ERCA/Cas12a) system for ultrasensitive microRNA detection. This novel method offers a simple, efficient, and highly specific approach for molecular diagnostics.
Area of Science:
- Molecular Biology
- Biotechnology
- Genetics
Background:
- MicroRNAs (miRNAs) are critical regulators of gene expression implicated in various diseases.
- Sensitive and specific detection of miRNAs is essential for molecular diagnostics and understanding disease mechanisms.
Purpose of the Study:
- To develop a novel ultrasensitive detection system for microRNAs.
- To combine exponential rolling-circle amplification with CRISPR/Cas12a technology for enhanced miRNA detection.
Main Methods:
- Development of a target-triggered exponential rolling-circle amplification (T-ERCA) system using a dumbbell probe.
- Integration of T-ERCA with CRISPR/Cas12a for signal amplification and specific target recognition.
- Validation of the T-ERCA/Cas12a system for miRNA detection in biological samples.
Main Results:
- The T-ERCA/Cas12a system demonstrated significantly higher amplification efficiency compared to single amplification methods.
- Ultrasensitive detection of miRNA-155 was achieved with a wide linear range (1 fM to 5 nM) and a low limit of detection (0.31 fM).
- The assay showed excellent specificity and applicability for assessing miRNA levels in different cell types.
Conclusions:
- The T-ERCA/Cas12a system provides a highly sensitive, specific, and simple method for miRNA detection.
- This approach holds significant potential for advancing molecular diagnostics and clinical applications.
- The developed system offers a new guidance for practical molecular diagnosis and clinical use.
